Phytochemistry and Pharmacological Activities of the Diterpenoids from the Genus Daphne.

There are abundant natural diterpenoids in the plants of the genus Daphne from the Thymelaeaceae family, featuring a 5/7/6-tricyclic ring system and usually with an orthoester group. So far, a total of 135 diterpenoids has been isolated from the species of the genus Daphne, which could be further classified into three main types according to the substitution pattern of ring A and oxygen-containing functions at ring B. A variety of studies have demonstrated that these compounds exert a wide range of bioactivities both in vitro and in vivo including anticancer, anti-inflammatory, anti-HIV, antifertility, neurotrophic, and cholesterol-lowering effects, which is reviewed herein. Meanwhile, the fascinating structure-activity relationship is also concluded in this review in the hope of providing an easy access to available information for the synthesis and optimization of efficient drugs.

Lathyrane diterpenoids from Jatropha podagrica and their antitumor activities in human osteosarcoma cells.

Two new lathyrane-type diterpenoids, jatropodagins A and B (1 and 2), and five known analogues (3-7), were isolated from the stems of Jatropha podagrica. Their structures and absolute configurations were elucidated by spectroscopic data and calculated ECD analyses. The cytotoxicities of all the lathyrane-type diterpenoids (1-7) were evaluated against two human osteosarcoma cell lines (Saos-2 and MG-63). Compound 1 exhibited significant cytotoxic effects against Saos-2 and MG-63 with IC50 values of 8.08 and 14.64 µM, respectively. The IC50 values for the positive control 5-FU against the Saos-2 and MG-63 cell lines were 19.01 and 25.00 µM, respectively. Morphological features of apoptosis activities were evaluated in 1-treated Saos-2 cells and the results confirmed apoptosis in a dose-dependent manner.

Oenothein B boosts antioxidant capacity and supports metabolic pathways that regulate antioxidant defense in Caenorhabditis elegans.

Oenothein B (OEB) has various biological functions, although few studies have focused on its effect on in vivo metabolic phenotypes. In the present study, the systematic antioxidant activity of OEB was evaluated both in vitro and in vivo, and the effect of OEB on metabolic pathways related to antioxidant capacity of Caenorhabditis elegans (C. elegans) was explored. Our findings indicate that OEB exhibits great antioxidant capacity and ability to scavenge free radicals and that OEB treatment can protect RAW 264.7 macrophages from oxidative damage by increasing superoxide dismutase (SOD) activity, catalase (CAT) activity and glutathione (GSH) content and the corresponding gene expression (sod2, cat, gpx1), while decreasing malonic dialdehyde (MDA) content. Moreover, OEB treatment significantly reduced ROS accumulation under oxidative stress conditions and increased glutathione peroxidase (GPx) activity and decreased MDA content in C. elegans. Metabolomics analysis revealed that sixteen out of forty-two significantly altered metabolites were selected as potential biomarkers related to alterations in the antioxidant status of worms, including metabolic pathways involved in amino acid metabolism, taurine and hypotaurine metabolism, lipid metabolism, and purine metabolism. Overall, our results provide new insights into the effects of OEB treatment on antioxidant capacity and metabolism that suggest that OEB could be a potentially good source of natural antioxidants.

Effects of resveratrol on the expression of molecules related to the mTOR signaling pathway in pathological scar fibroblasts.

BACKGROUND: We observed the effects of resveratrol on the expression of molecules involved in the mTOR signaling pathway in pathological scar fibroblasts, including PI3K, Akt and mTOR. METHODS: We detected the expression of PI3K, Akt and mTOR in pathological scar and normal skin fibroblasts through immunofluorescence. After being treated with different concentrations of resveratrol, the expression of PI3K, Akt and mTOR mRNA and protein were detected by RT-PCR and Western Blot respectively. RESULTS: Results showed that the expression of PI3K, Akt and mTOR were significantly enhanced in pathological scar fibroblasts and mainly expressed in the nucleus, with no expression in normal skin fibroblasts. Results from RT-PCR and Western Blot tests demonstrated that after Res intervention with different concentrations for pathological scar fibroblasts, the relative expression quantity of PI3K mRNA and protein decreased and showed a dose dependent relationship. Compared to the control group, the differences were statistically significant (P<0.05). However, decrease in the expression of PI3K mRNA and protein was not obvious and there were no significant differences in comparison to the control group (P>0.05). CONCLUSIONS: The mechanism of resveratrol in the inhibition of the proliferation of pathological scar fibroblasts may be related to its down-regulation in the expression of Akt and mTOR, which are the key molecules of mTOR signaling pathway.

Expression of mTOR/70S6K signaling pathway in pathological scar fibroblasts and the effects of resveratrol intervention.

The aim of the study was to examine the expression of mammalian target of rapamycin (mTOR)/70S6K signaling pathway in pathological scar fibroblasts and the effects of resveratrol (Res) intervention. The mTOR and 70S6K in pathological scar and normal skin fibroblasts were detected by immunofluorescence following treatment with different concentrations of Res. RT-PCR and western blot analysis were used to detect the expression of mTOR and 70S6K mRNA and protein, respectively. Immunofluorescence showed that the expression of 70S6K and mTOR was significantly enhanced in pathological scar fibroblasts, and mainly expressed in the nucleus, but not in normal skin fibroblasts. RT-PCR and western blot analysis showed that after different concentrations of Res treatments, the mTOR and 70S6K mRNA and protein expression significantly (P<0.05) decreased in a dose­dependent manner. In conclusion, the expression of mTOR/70S6K signaling pathway in pathological scar fibroblasts was significantly enhanced. Res can downregulate the expression of mTOR and 70S6K to achieve the inhibition of pathological scar fibroblast proliferation.

Expression of TGF-ß1/mTOR signaling pathway in pathological scar fibroblasts.

The aim of the present study was to detect the expression of the key molecules, including transforming growth factor­ß1 (TGF-ß1), phosphatidylinositol 3-kinase (PI3K), protein kinase B (Akt) of TGF­ß1/mammalian target of rapamycin (mTOR) pathway in pathological scar fibroblasts. Immunofluorescence, reverse transcription­polymerase chain reaction (RT­PCR) and western blot analysis were used to detect the expression of the key molecules TGF­ß1, PI3K, Akt, mTOR in fibroblasts of normal skin tissue and pathological scar tissue. Immunofluorescence showed that the expression of TGF­ß1, PI3K and Akt was significantly enhanced (P<0.05) in pathological scar fibroblasts, and mainly expressed in the cell nucleus, but not in normal skin tissue or fibroblasts. RT­PCR and western blot test results revealed that the TGF­ß1, PI3K, Akt, and mTOR mRNA and protein expression in pathological scar fibroblasts were significantly higher (P<0.05) than in the normal skin tissue. Expression of the TGF­ß1/mTOR signaling pathway in pathological scar fibroblasts was significantly increased. Data suggest that this expression may be an important mechanism for pathological scar formation.

Effect of Wubeizi ointment aqueous solution on the expression of type I and III procollagen genes in keloid fibroblasts.

We evaluated the effect of Wubeizi (WBZ) ointment on keloids. Keloid-derived fibroblast primary cultures were used to evaluate the effect of the different concentration of WBZ ointment on the expression of type I and III procollagen in keloid fibroblast primary cultures using dot blot assay. Type I and II precollagen cDNA probes labeled with non-radioactive digoxin were used for dot blot. Cell cultures were divided into 4 groups: The large dose group received 1 g/ml of WBZ, middle dose, and small dose groups received 0.5 and 0.25 g/ml of WBZ, respectively. The control group received serum-free medium without WBZ. Our results showed that type I and III procollagen mRNA expression was reduced significantly in the large dose and middle dose groups compared to the control group. Type I and III procollagen mRNA expression level in the small dose group had no statistically significant difference with the control group. However, the difference between the large dose group and the small dose group was statistically significant. We concluded that WBZ ointment aqueous solution restricted keloid fibroblast proliferation by downregulating the expression of type I and III procollagen and therefore reducing collagen deposition in keloid tissue.

Effects of resveratrol on the proliferation, apoptosis and telomerase ability of human A431 epidermoid carcinoma cells.

The aim of the present study was to investigate the effect of resveratrol on cell apoptosis, ability of telomerase and the human telomerase reverse transcriptase (hTERT) protein expression in human A431 epidermoid carcinoma cells. A431 cells were treated with different concentrations of resveratrol, and the cell appearance was then observed under a microscope. In addition, the cell proliferation was examined using an MTT assay, and the ability of telomerase was detected using telomeric repeat amplification protocol-polymerase chain reaction-ELISA. Resveratrol significantly inhibited the ability of telomerase and decreased the expression of hTERT protein in a concentration-dependent manner. In conclusion, resveratrol is capable of downregulating the expression of hTERT protein and inhibits the ability of telomerase of A431, which is an important mechanism of action of resveratrol with regard to inhibition of A431 cell proliferation.

Resveratrol Inhibits Proliferation and Induces Apoptosis of Pathological Scar Fibroblasts Through the Mechanism Involving TGF-ß1/Smads Signaling Pathway.

The objective of this study was to determine the effect of resveratrol (Res) treatment on pathological scar fibroblasts and the changes in TGF-ß1/Smads signaling pathway. For this purpose, cultured pathological scar fibroblasts were treated with various concentrations of Res (10, 50, and, 100 µmol/l), and the morphological changes in target cells were studied using scanning electron microscopy (SEM). The cellular proliferation was assessed by MTT assay; the mRNA and protein expressions of TGF-ß1 and Smad-2,3,4,7 were determined by reverse transcription polymerase chain reaction (RT-PCR) and immunofluorescence assay, respectively. We found that Res-treated fibroblasts exhibited the typical apoptotic morphological changes. As shown by MTT assay, the OD values of Res-treated fibroblasts, as a measure of cell growth, were significantly lower than those of controls (P < 0.05). In addition, as compared to controls, TGF-ß1 and Smad-2,3,4 mRNA/protein expression decreased but those of Smad7 increased in a dose-dependent manner (P < 0.05). It was, therefore, concluded that Res treatment inhibited the pathological scar fibroblast proliferation and induced cell apoptosis through the mechanism involving downregulation of TGF-ß1, Smad-2,3,4, and upregulation of Smad7.

The effects of Wubeizi ointment on the proliferation of keloid-derived fibroblasts.

UNLABELLED: To evaluate the effectiveness of the Wubeizi (WBZ) ointment on keloid-derived fibroblasts. The primary cells of the keloid-derived fibroblasts were cultured and the effectiveness of the WBZ ointment at different concentrations was examined by MTT colorimetric methods on keloid-derived fibroblasts. The WBZ ointment showed inhibitory effects on proliferating the keloid-derived fibroblasts (P < 0.01)in a time- and dose-dependent manner. The proportion of cells in S stage was significantly higher in each of the WBZ ointment group than in the control group (P<0.01), and the proportion of G2 + M stage cells was significantly lower than that of control group, which was statistically significant (P < 0.01).The inhibitory effects of the S and G2 + M stage increased with higher drug concentrations (P < 0.05). CONCLUSION: The WBZ ointment can inhibit the proliferation of the keloid-derived fibroblasts in a time- and dose- dependent manner.